Antibody Panel to Green Fluorescent Protein (GFP) - FocusOn 118

Green Fluorescent Protein (GFP), due to the property of exhibiting bright green fluorescence when exposed to blue or UV light, has emerged as a powerful research tool for assessing gene expression and subcellular protein distribution in fixed or living cells. This feature makes it ideal as a marker for use in fluorescence microscopy, cytometry, tagging fusion proteins, and assaying transcriptional regulation from gene promoters in vivo.

Appealing in its ease of use, GFP folds into a chromophore without the need for accessory cofactors, enzymes or substrates other than molecular oxygen. Originally isolated from the jellyfish A. victoria, a number of laboratory mutations have been engineered into GFP to produce experimentally desirable variants with increased fluorescence, photostability, folding efficiency, and different excitation and emission peaks (blue, green, and yellow). The GFP gene or its variants are frequently attached to genes of interest using standard molecular and cell biology techniques to produce chimeric proteins, wherein the GFP serves as a reporter on the localization of the target protein. The small size and well-folded beta barrel structure make the highly fluorescing GFP relatively inert to the structure and properties of target proteins, and therefore ideal for this application. The GFP gene (as well as variants isolated from other marine species or engineered through mutations) has been introduced and expressed in a diverse set of model systems in bacteria, yeast and other fungi, fish, plants, insects, and mammalian cells and organisms.

Acris Antibodies offers a wide range of antibodies to detect GFP and its variants in various applications. The table below shows our recommended antibodies.

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Fig. 1: Confocal microscopy images of COS-7 cells transfected with expression constructs encoding membrane-tethered EGFP (membrane-EGFP; top) or nuclear Polycomb 2-EYFP fusion protein (Pc2-EYFP; bottom). The natural fluorescence of the produced proteins is shown in the green channel (left), the anti-GFP antibody Cat.-No. SP3005P signal was detected in the red channel (right). The system was carefully tested for overlap of these two optical channels and images were scanned separately in sequential scanning mode. The blue nuclear stain is also shown.

Fig. 2: Immunoprecipitation of GFP-NLS from HEK293 cells lysed in non-denaturating conditions using a rabbit anti-GFP antibody (lane 2) or a pre-immune rabbit serum (lane 3). Immunoprecipitates together with a sample of the cell lysate (lane 1) were separated on SDS-PAGE polyacrylamide gel and immunoblotted with the anti-GFP antibody Cat.-No. SP3005P.